Dataset Information

Burkholderia cenocepacia K56-2 transcriptomic profile during the early contacts with surface of host bronchial epithelial cells

ABSTRACT: In this work, high-throughput RNA sequencing was used to assess transcriptomic alteration in Burkholderia cenocepacia K56-2 in the early stages of interaction with host-cell surface. Giant plasma membrane vesicles (GPMVs) from 16HBE14o- bronchial epithelial cell line were produced and used as a cell-like alternative. RNAseq was performed and the expression of the adherent (GPMV-attached) and non-adherent (planktonic control) bacteria were compared. Using a fold change cut off of ≥ 1.5 (adjusted P-value < 0.01), the obtained RNAseq dataset indicates a total of 926 genes which expression was altered upon bacteria-GPMV contact. The sensing of host membranes lead to a transcriptional shift that cause a cascade of metabolic and physiological adaptations to the host specific environment. Many of the differentially expressed genes encode proteins related with central metabolic pathways, transport systems, cellular processes, and virulence traits.

ORGANISM(S): Burkholderia cenocepacia

PROVIDER: GSE155982 | GEO | 2021/03/01

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:BACKGROUND: We identified that a putative CzcRS-like two-component system (TCS) in Burkholderia cenocepacia K56-2 is required for heavy metal resistance and virulence. In an attempt to identify genes directly regulated by the CzcR response regulator, we performed ChIP-seq analysis to identify genomic regions bound by CzcR. METHODS. Sequence encoding a FLAG octapeptide (Sigma-Aldrich) was introduced to the 3’ end of the czcR gene at its native chromosomal position in wild-type B. cenocepacia K56-2. Wildtype K56-2 and the CzcR-FLAG strain were grown in 50 ml tryptone soya broth (TSB) in the presence or absence of 1.5 mM zinc chloride. After overnight incubation, anti-FLAG immunoprecipitation of CzcR-bound genomic DNA was performed. Wildtype K56-2 was processed in parallel, to serve as a control against which to assess for enrichment of genomic regions. Illumina sequencing libraries were prepared from the resulting DNA using the Nextflex ChIPseq protocol (Bioo Scientific) with indexed adapters. Libraries were amplified by 18 cycles PCR, purified using 0.8 volumes Ampure XP beads (Beckman Coulter) and quantified with a Bioanalyzer 7500 assay (Agilent). Libraries ranged in size from 150 bp to 800 bp with a average insert size of 310 bp. Libraries were pooled in equimolar concentrations, denatured and diluted to 6.5 pM, clustered on a flowcell using a cBOT (Illumina) and sequenced on a HiSeq2000 (Illumina). Paired-end reads were filtered using the fastq-mcf package from the ea-utils suite to remove reads with less than 90% Q20 scores or above and to trim off adaptor sequence. The reads were then aligned against the B. cenocepacia J2315 genome (NC_011001-NC_011003) using BWA (0.5.9) and converted to BAM format using Samtools. Potential PCR duplicates were removed using the samtools rmdup command. The MACS package (v1.4) was used to compare and contrast the control and sample data using the –call-subpeaks and –w options. RESULTS: Relative to wildtype B. cenocepacia K56-2, the only genomic region that was found to be enriched by the ChIP-seq analysis of the CzcR-FLAG strain mapped to nucleotide coordinates 787809-798988 on chromosome 2. This region includes the czcRS genes and those encoding the associated efflux pump (CzcCBA).

Dataset Information

Burkholderia cenocepacia K56-2 transcriptomic profile during the early contacts with surface of host bronchial epithelial cells

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets